CN1068161A

CN1068161A - Method for delignification and bleaching of lignocellulose-containing material and wastewater treatment with synergistic laccase

Info

Publication number: CN1068161A
Application number: CN92104549.2A
Authority: CN
Inventors: 汉斯-彼得·考尔
Original assignee: Individual
Current assignee: Individual
Priority date: 1991-05-17
Filing date: 1992-05-16
Publication date: 1993-01-20
Also published as: NO934148D0; BR9206013A; DE4137761A1; FI935067L; WO1992020857A1; JPH06508663A; FI935067A7; FI935067A0; AU1793392A; NO934148L; AU663501B2; CA2103260A1; EP0584176A1

Abstract

The invention relates to a method for degrading lignin in lignocellulose-containing material and/or bleaching lignocellulose-containing material in order to enzymatically refine old paper pulp and treat paper or pulp mill waste water, wherein laccase from Coriolus versicolor is used, and simultaneously and/or quantitatively non-aromatic reducing and oxidizing agents and phenolic compounds and/or non-phenolic aromatics are added, the latter two substances directly oxidizing the phenolic and non-phenolic structures of lignin by the action of these phenolic and/or non-phenolic aromatics or oxidizing lignin by means of oxidation reactions of other phenolic and/or non-phenolic compounds caused by these compounds.

Description

Lignocellulose-containing materials delignification, bleaching and with the method for the treatment of waste water by synergetic shellac enzyme

The present invention relates to a kind ofly make lignocellulose-containing materials delignification and/or bleaching so that old paper paper pulp enzyme is urged refine and handle the especially novel method of papermaking or pulp mill wastewater of waste water.

The prior art of I, pulp preparation chemical method, the application of wooden lyase: bio-pulping, other application

The chemical method that A, pulp preparation and lignin produce

Nowadays being used for the pulp preparation main method is sulfate process and sulphite process.These two kinds of methods all are that boiling produces paper pulp under pressure.Sulfate process is used effective chemicals NaOH and Na ₂The S operation, sulphite rule Ca(HSO ₃) ₂+ SO ₂

In addition, also there are several boiling methods that alleviate environmental pollution by means of organic solvent.

The main target of all methods is: to remove lignin in used vegetable material timber or the annual plant.Lignin has been formed the main component of vegetable material (stem or trunk) with cellulose and hemicellulose, lignin must be removed, otherwise just can not make the paper of not flavescence and anti-high load capacity.

These pulp preparation methods are utilized stone to grind (groundwood pulp) or are operated with refiner (TMP), and they make the timber through appropriate pretreatment (chemistry, heat or thermochemical method) grind into wood pulp by corresponding grinding.

This paper pulp contains lignin greatly, is used for making newspaper printing or printing magazine or advertisement paper, is commonly referred to as coated paper here.

B, bio-pulping

In recent years, just the begin one's study mechanism of action of wooden dissolution system of people was recognized to plant in the mould at white rot can make golden spore Pseudomonas Phanerochaete produce the enzyme of q.s by suitable condition of culture and inductor additive.The enzyme class of finding in this mould is: lignin peroxidase and manganese dioxide enzyme, their this form past is unknown.Because this mould is a kind of efficient lignin degradation body in vivo, thereby at first assert, also can xylogen degradation with no fungous enzyme the time.It is wrong that this imagination is proved to be, because this kind of enzyme also causes the reunion rather than the degraded of lignin.

The peroxidase of other dissolved lignin and the enzyme of other dissolved lignin such as laccase also similarly, they use O ₂Replace H ₂O ₂Come the oxidative degradation lignin.In either case, degraded is all carried out with similar process, promptly forms a kind of atomic group, if these atomic groups are not hunted down in any mode, they be can do by myself and react to each other again and regroup.Therefore, people have to when biological pulp it be transformed system in the ex vivo.

People's cracking mould and for example wood chip, so that partly remove lignin, this can improve bleaching property at least, and reduces the required energy of mechanical treatment (defiber) after this.Bio-pulping be generally understood as with biology enzyme urge legal system be equipped with paper pulp or with material like the pulp type or pre-product.Bio-pulping also will be tested earlier.Cultivate the bacterial strain that does not form cellulase, promptly so-called cellulose-less enzyme mutant body, reason is only to wish to obtain the lignin lyase, forms cellulase even this this enzyme also can make white rot plant mould in the presence of ligno-ccllulose.Thereby cellulase may cause undesirable productive rate to reduce owing to cause cellulose degradation in paper pulp.

Another kind of difficulty is that white rot is planted mould and can not therefore be transferred to make carbon source with cellulose or hemicellulose with lignin as carbon source.At present handle these difficulties, promptly flood matrix in addition, so that cellulose-less enzyme mutant body might be grown with a kind of additional carbon source such as glucose with following method.

Here, main difficulty is lignin degradation speed very slow (several days to several weeks), therefore in vivo in the method: make wood fragments consider ten cellulose-less enzyme mutant bodies (golden spore Pseudomonas Phanerochaete) to be worth doing and process by sieving new better wild strain.As final goal, people try hard to faster and lignin degrading effectively, with the purpose of being mentioned above reaching: improve grind can reduction and the raising of bleaching property.

If can successfully make the loss of cellulose and hemicellulose keep very little, people will obtain the high yield material, but opposite with the groundwood pulp of grinding preparation up to now, it can prepare with extremely low energy consumption, and will present easier bleaching and better mechanical performance.

As previously mentioned, Sheng Xia major defect is:

1, the degraded of this element is very slow;

2, lignin degradation very limited (low %)

3, it seems and to add additional C source inevitably;

4, because growth and degradation time are long, need to ventilate especially, keep PH constant and must not have impurity, cause very big deposition volume, thereby be the difficulty of bringing of process.

Also there is deposit during with acellular enzyme system and golden spore Pseudomonas Phanerochaete.

Yet, the application here is: 1) " kraft pulp " (promptly part is sloughed the paper pulp of lignin, it forms the residual lignin content of about 5-8% behind sulphate cook), 2) improve the mechanical performance of mechanical pulp, just refer to wood pulp is carried out the paper pulp that forms after machinery-Re or chemical-mechanical or the chemistry-thermal-mechanical treatment, TMP(thermal-mechanical treatment paper pulp for example), CMP(chemical-mechanical pulp treatment) and CTMP(chemistry-thermal-mechanical treatment paper pulp) and 3) decolouring (referring to the paper pulp that kraft pulp-level bleaching back forms) of E1 waste water.

The application here is limited to degraded to low percentage (wood pulp) lignin percentage from (" kraft pulp ") in 12 and 48 hours.Bleach unsatisfactory equally.

People have done test several times in the recent period, utilize blood thing (Haemverbindungen) to obtain better bleaching results as hemoglobin, and these materials are regulated the active blood component (Haemkomponente) in the lignin peroxidase to a certain extent.These compounds are normally too expensive for commercial Application, and partly have undesirable contingent effects, like cellulosic degraded.

A kind of enzyme of other type, hemicellulase refers to zytase here, also as bleaching enzymes, reason is that people estimate the influence that is linked with between hemicellulose (xylan) and the lignin.

Here the long action time of enzyme, discoloration is little and can not separating lignin, has significant disadvantages equally.

Other application of wooden lyase is the decolouring of pulp manufacturer waste water and the oxidation and the detoxification of detoxification and chlorination aromatic substance and other compound.Also not having gratifying solution, reason at present in this field is that the test of the decolouring in the mould fungus strain and detoxification all use freely or the mould of fixing carries out in body up to now.Have only through the time (several days) of length just can reach gratifying degraded or oxidation number result.

The task that the present invention proposes is, lignin in the exploitation degraded lignocellulose-containing materials and/or bleaching lignocellulose-containing materials and enzyme are urged and refined old paper paper pulp and handle the method for papermaking and paper plant's waste water, and this method should not have above-mentioned shortcoming.

This task can be thus completed: use the laccase that is obtained by variegated aspergillus straw mulch Pseudomonas (Coriolus Versicolor), add simultaneously or allocate non-fragrant reductant and oxidant and those oxybenzene compounds and/or non-phenol aromatic into, they are by the phenol or the non-phenol structure of the effect direct oxidation lignin of phenol or non-phenol aromatic, perhaps by means of finish by these compounds other phenol and/or the oxidation of non-phenol compound come further oxidative lignin.

German patent application DE 3636208(WO88/03190), P4088 93.6, though P408894.4 has described the application of wooden lyase, also added reductant, oxidant and other reducing/oxidizing material (salt etc.) simultaneously.

But with the main distinction of the document be to relax to have used laccase, that is to say, by adding specific material (phenol aromatic substance, non-phenol aromatic substance) obviously increased the laccase of its validity, thereby can corrode whole lignin molecule, work as a kind of lignin peroxidase is the sort of.

The present invention uses the laccase of variegated aspergillus Coriolus Qu61.This laccase adds with laccase base-material (phenol compound or non-phenol fragrance material) and/or oxybenzene compound and/or non-phenol aromatic substance, these compounds are oxidized, thereafter they perhaps carry out oxidation by other reducing/oxidizing intermediate (oxybenzene compound or non-phenol aromatic substance such as veratryl alcohol) that adds therebetween as reducing/oxidizing intermediate direct oxidation lignin.So, the laccase that only corroded the phenol component (only account for its separation structure about 1/3) of lignin also can be invaded only non-phenol separation structure, like this, can cause the degraded fully of whole lignin molecule in theory, just as a kind of lignin peroxidase, work fully.

Wherein, these reducing/oxidizing systems must " be fit to " mutually, just as the already oxidised material in the respiratory chain is done the oxidant effect of other material, and are reduced by this material.Fig. 1 represents that laccase is at prothetic group (Cu ²⁺Cu ¹⁺) action principle on the plane.

Fig. 1

2(Cu ²⁺) laccase+phenol

2(Cu ⁺) laccase+atomic group (I)+2H ⁺

2(Cu ⁺) laccase+Y ₂O ₂+ 2H ⁺

2(Cu ²⁺) laccase+H ₂O.

Laccase can impel phenol to produce a kind of mesomer aryloxy group.This is a kind of new deposit.

Opposite with the method for the described use of above-mentioned patent laccase, a kind of major advantage is, the cultivation of variegated aspergillus Coriolus Qu61 is quite simple, and lignin peroxidase culture such as golden spore Pseudomonas Phanerochaete then will cultivate in bigger round very difficultly.The productive rate of itself is about 20Miu IU/l with a kind of special inductor the time.Because the effect of laccase has expanded the similar characteristic of lignin peroxidase to, thereby might realize the big industrial applications of this kind of enzyme, this is a tangible advantage.Only use laccase (that is to say and do not have above-mentioned system) and make base-material with pine tree/wood pulp with laccase and phenol of between this quality and enzyme, changing or non-phenol reducing/oxidizing intermediate composition, be about at initial content of lignin under 27% the situation and carry out the lignin degradation test, lignin can be remained content at most and be removed to 22%, just degradation rate is about 19%.Can make the lignin residual content be reduced to 16% with new system, degradation rate is about 41%, and it is many to be original twice.

During processing, used temperature range is 25-60 ℃, and pH regulator is to 3-8.

By adding reductant and/or oxidant, salt and/or oxybenzene compound, the present invention can access the redox potential of 200-600mv.This available in the methods of the invention oxidation-reduction electrode records, and regulates with an adjuster and control device, and total reaction time remains unchanged by adding Oxidizing and Reducing Agents, salt and oxybenzene compound.

Preferentially select air, O for use as oxidant ₂Or hydrogen peroxide.Or organic peroxide.Can consider sodium hydrogensulfite, ascorbic acid, dithionite and sulfur-containing compound or the sweet peptide of wheat protein as reductant.

As the salt slaine.Here especially consider FeSO ₄, FeCl ₃, MnSO ₄, TiO ₂, CuSO ₄And Mn ₃-acetic acid and waterglass.Be also pointed out that in addition the present invention preferably adds complexing agent.Complexing agent can be used for example ethylenediamine tetra-acetic acid (EDTA) or diethylene-triamine pentaacetic acid (DTPA).

Except that above-mentioned substance, pulp-water solution also can contain other material.Admissible have NaOCl, washing agent and a polysaccharide.Described polysaccharide comprises that glucan, mannosan, dextran, Luevan or vegetable rubber and/or self are formed by mould or is mixing the polysaccharide that produces when cultivating with yeast.Aliphatic acid of also available nonionic, ion, anion, CATION and amphoteric surfactant, soap class such as saponification and glucan and xanthan alkane (Xanthane) to this.In addition, except wooden lyase, also can assistance application hemicellulase and/or pectase and/or lipase.

With article " oxidation of non-phenol base-material " (267 phases of FEBS of R.Bourbonnais and M.G.Paice, nineteen ninety, the 99-102 page or leaf) main distinction of comparing is the embodiment difference of process, promptly add reductant and oxidant simultaneously, promptly add the atomic group component and add the atomic group acceptor, so that make the depolymerization of lignin preferential, and when keeping reaction, oxidant suppressing to meet again by adding by " catching atomic group ".Known laccase also with H ₂O ₂Concur, in certain base-material, peroxide can be strengthened the oxidation (R.Blaich, Analytische Elekt-rophoreseverfahren, 1978,59 pages of Thieme publishing houses) of laccase.

System of the present invention can be used laccase separately, also can plant mould with lignin peroxidase and manganese dioxide enzyme for example golden spore Pseudomonas Phanerochaete or other white rot and use.The latter, promptly lignin peroxidase (manganese dioxide enzyme)+laccase is to operate under the condition of 4-5 at PH, at this moment two fermentoids all are active.Real lignin degradation, bleaching (with the lignin degradation while) or enzyme urge the expansion process of refining by adding reductant and oxidant simultaneously, promptly cause by the atomic group of catching lignin degradation formation.The Fe of special metal salt such as NaOCl ²⁺, Fe ³⁺, Cu ²⁺, Ti ³⁺The time to exist usually be important, their produce specific atomic group as a ° OH-group (Fe ^{+ 2}And/or Ti ³⁺+ H ₂O ₂), ° O for example ₂H-group (Ti ³⁺And O ₂) or Sigulett oxygen (NaOCl+H ₂O ₂=NaCl+H ₂O+O ₂), these groups can be accelerated lignin degradation.Important auxiliary agent is the existence of polysaccharide and the existence of washing agent, polysaccharide is now by mould generation itself or from yeast cell wall, the latter plants to mix between mould and the yeast when cultivating at white rot and forms, washing agent can make enzyme penetrate between the fiber better in delignification and/or bleaching process, and foaming and collecting action for ink particles when deinking are absolutely necessary.When deinking, a kind of additional bleach system may be a no less important as sulfinic acid carbonamidine (FAS)/NaOH, so that the NaOH/H that the compensation chemical system exists ₂O ₂Discoloration, soap class additive itself or combine with washing agent and also can be used as battery.

When enzyme is urged pulp treatment waste water (TMP/ wood pulp etc.), use laccase+phenol or non-phenol aromatic substance.Here,, do not add reductant, add reductant (H continuously but have by on purpose adding phenol or the non-phenol aromatic substance of determining ₂O ₂, O ₂Or air oxygen), might make the almost completely polymerization of lignin that contains in the waste water, than single with the value that laccase reached high (20-50%).Thereby might be by becoming soluble and the lignin of 70-90% is removed in flocculation thus, sedimentation, filtration, little flotation etc. from waste water, and astoundingly CSB be reduced to 60-70%.

A, embodiment 1)

Embodiment: the enzyme of sulfate pulp is urged bleaching

The absolutely dry paper pulp of 50g (sulfate pulp) is stirred under 1% pulp thickness, 40 ℃ and about 500rpm condition in a stirred vessel.With 1n HCl pH value is transferred to 5.Adding 0.1-1.5%(is benchmark with the adiabatic drying material) H ₂O ₂, about 0.1% ETDA or DTPA, 0.01-0.5%(be benchmark with absolute dry material) copper sulphate (II).Add 500-8000IU(1Iu=1 μ m syringaldazine transform/minute/the ml enzyme) behind the laccase of variegated aspergillus Coriolus Qu61, add about 2 * 10 by the adiabatic drying material ^-3%-1X10 ^-10% veratryl alcohol and 0.01-0.1% naphthols are again by being metered into H simultaneously ₂O ₂Begin bleaching process with solution of sodium bisulfite.Make redox potential be maintained at about 400mV.After this process begins, made it to continue 2-4 hour.Carry out the control and the adjusting of technology with an oxidation-reduction electrode and a pump control device.

C, embodiment 2)

The delignification of paper pulp (TMP)

It is 5 that the solution (1%) that the definitely dried TMP of 100ml water and 1g is formed with 1m HCl is adjusted to pH value.Add 8 μ mol veratryl alcohols, 400 μ mol H by the absolute dry material of every g ₂O ₂With the 0.01-0.1% naphthols.Add sodium hydrogensulfite and H in batches simultaneously (1ml/ hour) ₂O ₂Solution (each 9.80mmol).The 100000Iu enzyme (laccase of variegated aspergillus Coriolus Qu61) that adds makes the reaction beginning.Be reflected under 40 ℃ and the about 500rpm stirring condition and carried out 4 hours.

D, embodiment 3)

Embodiment: have pretreated enzyme to urge deinking

The old paper of 100g over dry (50% newspaper/50% magazine) is torn into the big fragment of 1 * 2cm.Water 2l water (about 45 ℃) to it, pH value is transferred to 7-7.5 with alum.Then by the old paper of every g over dry, add 80-8000IU(1IU=1 μ mol syringaldazine transform/minute/the ml enzyme) laccase, 8 μ mol veratryl alcohols and 0.05-0.6% washing agent make battery and 0.2-2 μ l solution of sodium bisulfite (38-40%).In the disintegrator of 3000rpm, collided 15 minutes then.Old paper pulp was placed 75 minutes at about 45 ℃, or in the industrial flotation cell of a 20l flotation 10-20 minute immediately.

Embodiment 4)

The enzyme of wood pulp waste water (TMP. groundwood pulp etc.) is urged processing〉〉〉polymerization of the lignin that exists.

With every liter of 10-200000IU laccase+0.01 * 10 ^-3-0.1% the unfiltered TMP waste water of naphthols dilution 250m makes it to cultivate 1-4 hour in 20-50 ℃ under the condition of stirring and bubbling air oxygen.The polymerization that obviously improves with laccase than single has taken place in the lignin that exists, and become soluble thing, and can be removed by adding suitable flocculant such as polymine, or by the micro-filtration centrifugation or remove by filter.

Claims

1, degraded in the lignocellulose-containing materials lignin and/or the bleaching lignocellulose-containing materials so that enzyme is urged refines old paper paper pulp and handle papermaking and the method for pulp mill wastewater, it is characterized in that, use forms the laccase that thing obtains by variegated aspergillus Coriolus Qu61 and other laccase, add and/or quantitatively add non-aromatics reductant and oxidant and following phenol compound and/or non-phenol aromatic substance simultaneously, the phenol and the non-phenol structure of this two classes material or the effect direct oxidation lignin by phenol and/or non-phenol aromatic substance perhaps come oxidative lignin by means of other phenol that is caused by these compounds and/or the oxidation reaction of non-oxybenzene compound.

2, by the method for claim 1, it is characterized in that, except that laccase, auxiliary lignin peroxidase and/or the manganese dioxide enzyme of adding.

3, by the method for claim 2, it is characterized in that, use by golden spore Pseudomonas Phanerochaete or other white rot and plant lignin peroxidase and/or the manganese dioxide enzyme that mould produces.

4, by the method one of in the claim 1 to 3, it is characterized in that, add hemicellulase and/or pectase and/or lipase to reaction solution in addition.

5, by the method one of in the claim 1 to 4, it is characterized in that, pH value is adjusted between 3 to 8.

6, by the method one of in the claim 1 to 5, it is characterized in that temperature is between 25 to 60 ℃.

By the method one of in the claim 1 to 6, it is characterized in that 7, redox potential is 200 to 600mv.

8, by the method for claim 7, it is characterized in that, in reaction solution, add slaine in addition.

9, by the method for claim 8, it is characterized in that,, use FeSO as slaine ₄, FeCl ₃, MnSO ₄, TiO ₂, CuSO ₄And Mn ₃-acetate.

10, by the method for claim 9, it is characterized in that, add NaOCl.

11, by the method one of in the claim 8 to 10, it is characterized in that, make reductant with sodium hydrogensulfite, sodium dithionate, ascorbic acid, sulfur-containing compound or the sweet peptide of wheat protein.

12, by the method one of in the claim 8 to 11, it is characterized in that, with air, oxygen, H ₂O ₂Or organic peroxide is made oxidant.

13, by the method one of in the claim 1 to 12, it is characterized in that, also add washing agent in addition.

14, by the method for claim 13, it is characterized in that, make washing agent with surface-activitys nonionic, ion, anion, CATION and both sexes.

15, by the method one of in the claim 1 to 14, it is characterized in that, in reaction solution, add polysaccharide in addition.

16, by the method for claim 15, it is characterized in that, add glucan, mannosan, dextran, Laevan or vegetable rubber and/or self forms or the polysaccharide of generation when mixing cultivation with yeast by mould as polysaccharide.

17, by the method for one of claim 1 to 16, it is characterized in that, in reaction solution, add complexing agent in addition.

18, by the method for claim 17, it is characterized in that, make complexing agent with ethylenediamine tetra-acetic acid (EDTA) or diethylene-triamine pentaacetic acid (DTPA).

19, by the method for claim 1 to 18, it is characterized in that, add waterglass.

20, by the method for claim 1 to 19, it is characterized in that, add for example aliphatic acid of saponification of soap class.

21, by the method for claim 1 to 20, it is characterized in that, add formamidine sulfinic acid (FAS)/NaOH to this system.

22, by the method for claim 1 to 21, it is characterized in that, add the atomic group component to this system.

23, by the method for claim 1 to 22, it is characterized in that, add the atomic group acceptor to this system.

24, by the method for claim 1 to 23, it is characterized in that, be metered into laccase, lignin peroxidase, manganese dioxide enzyme or their mixture as enzyme.